title: Structural stability of Burkholderia cenocepacia biofilms is reliant on eDNA structure and presence of a bacterial nucleic acid binding protein.
abstract: Cystic fibrosis (CF) is the most common lethal inherited genetic disorder affection Caucasians. Even with medical advances, CF is life-shortening with patients typically surviving only to age 38. Infection of the CF lung by Burkholderia cenocepacia presents exceptional challenges to medical management of these patients as clinically this microbe is resistant to virtually all antibiotics, is highly transmissible and infection of CF patients with this microbe renders them ineligible for lung transplant, often the last lifesaving option. Here we have targeted two abundant components of the B. cenocepacia biofilm for immune intervention: extracellular DNA and DNABII proteins, the latter of which are bacterial nucleic acid binding proteins. Treatment of B. cenocepacia biofilms with antiserum directed at one of these DNABII proteins (integration host factor or IHF) resulted in significant disruption of the biofilm. Moreover, when anti-IHF mediated destabilization of a B. cenocepacia biofilm was combined with exposure to traditional antibiotics, B. cenocepacia resident within the biofilm and thereby typically highly resistant to the action of antibiotics, were now rendered susceptible to killing. Pre-incubation of B. cenocepacia with anti-IHF serum prior to exposure to murine CF macrophages, which are normally unable to effectively degrade ingested B. cenocepacia, resulted in a statistically significant increase in killing of phagocytized B. cenocepacia. Collectively, these findings support further development of strategies that target DNABII proteins as a novel approach for treatment of CF patients, particularly those whose lungs are infected with B. cenocepacia.
authors.affiliation: Center for Microbial Pathogenesis, The Research Institute at Nationwide Children's Hospital, and The Ohio State University College of Medicine, Columbus, Ohio, United States of America.
authors.forename: Laura A
authors.initials: LA
authors.lastname: Novotny
authors.forename: Amal O
authors.initials: AO
authors.lastname: Amer
authors.forename: M Elizabeth
authors.initials: ME
authors.lastname: Brockson
authors.forename: Steven D
authors.initials: SD
authors.lastname: Goodman
authors.forename: Lauren O
authors.initials: LO
authors.lastname: Bakaletz
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D000907:Antibodies, Bacterial / Q000502:physiology; D001426:Bacterial Proteins / Q000276:immunology*; D001483:Base Sequence; D018441:Biofilms; D057508:Burkholderia cenocepacia / Q000187:drug effects / Q000502:physiology*; D002460:Cell Line; D016384:Consensus Sequence; D003550:Cystic Fibrosis; D004269:DNA, Bacterial / Q000235:genetics*; D004268:DNA-Binding Proteins / Q000276:immunology*; D005110:Extracellular Space / Q000235:genetics; D006801:Humans; D007106:Immune Sera / Q000502:physiology; D034982:Integration Host Factors / Q000737:chemistry / Q000276:immunology; D016676:Macrophages, Alveolar / Q000276:immunology / Q000382:microbiology; D051379:Mice; D050296:Microbial Viability / Q000187:drug effects / Q000276:immunology; D008969:Molecular Sequence Data; D011817:Rabbits; D013183:Sputum / Q000382:microbiology
chemical_list: D000900:Anti-Bacterial Agents; D000907:Antibodies, Bacterial; D001426:Bacterial Proteins; D004269:DNA, Bacterial; D004268:DNA-Binding Proteins; D007106:Immune Sera; D034982:Integration Host Factors